Description
Product details:
HRP Conjugation Kit / HRP Labeling Kit (ab102890) uses a simple and quick process for HRP labeling / conjugation of antibodies. It can also be used to conjugate other proteins or peptides.
To conjugate an antibody to HRP using this kit:
- add modifier to antibody and incubate for 3 hrs
- add quencher and incubate for 30 mins
The HRP conjugated antibody can be used immediately in WB, ELISA, IHC etc. No further purification is required and 100% of the antibody is recovered for use. Learn about buffer compatibility below . Custom size conjugation kits up to 100 mg are available on demand. Please contact us to discuss your requirements.
This product is manufactured by Expedeon, an Abcam company, and was previously called Lightning-Link
HRP Labeling Kit. 701-0004 is the same as the 5 mg size. 701-0003 is the same as the 5 x 1 mg size. 701-0000 is the same as the 3 x 100 ug size. 701-0030 is the same as the 3 x 10 ug size. 701-0010 is the same as the 100 μg size. 701-0002 is the same as the 1 mg size.
Amount and volume of antibody for conjugation to HRP
Kit size
Recommended
amount of antibody
Maximum
amount of antibody
Maximum antibody
volume
3 x 10 μg
3 x 10 μg
3 x 40 μg
3 x 10 μL
100 μg
1 x 100 μg
1 x 400 μg
1 x 100 μL
3 x 100 μg
3 x 100 μg
3 x 400 μg
3 x 100 μL
1 x 1 mg
1 x 4 mg
1 x 1 mL
5 x 1 mg
5 x 1 mg
5 x 4 mg
5 x 1 mL
1 x 5 mg
1 x 20 mg
1 x 5 mL
Recommended amount of antibody will give an HRP:antibody molar ratio of 4:1. Antibodies often also perform well at the maximum amount of antibody which is 1:1 molar ratio.
Ideal antibody concentration is 1mg/ml. 0.5 - 1 mg/ml can be used if the maximum antibody volume is not exceeded. Antibodies > 4 mg/ml or < 0.5 mg/ml should be diluted /concentrated.
Buffer Requirements for Conjugation
Buffer should be pH 6.5-8.5.
Buffer constituent compatibility
If a concentration is shown, then the constituent should be no more than the concentration shown. If several constituents are close to the limit of acceptable concentration, then this can inhibit conjugation.
Compatible buffer constituents
50mM / 0.6% Tris
0.1% BSA
50% glycerol
0.1% sodium azide
Potassium phosphate
Sodium chloride
Sucrose
Sodium citrate
Trehalose
Tris buffered saline is almost always ≤ 50 mM / 0.6%
Sodium azide irreversibly inhibits HRP; antibodies with azide should be purified before using the HRP conjugation kit.
Incompatible buffer constituents
Thiomerosal
Proclin
Glycine
Arginine
Glutathione
If a constituent of the buffer containing your antibody or protein is not listed above, please contact us.
Only purified antibodies are suitable for use, ie. where other proteins, peptides, or amino acids are not present: antibodies in ascites fluid, serum or hybridoma culture media are incompatible.
Storing and handling conjugation kits
Lyophilized Lightning-Link
components are hygroscopic.
Kits are intentionally shipped at ambient temperature with silica gel to avoid exposure to moisture. Upon receipt, store the kit frozen and protect from moisture. Before opening the outer container, allow the lyophilized components to reach room temperature to minimize condensation.
Properties and Storage Information:
Shipped at conditions-Ambient - Cannot Ship with Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
